A fluorescent derivative of methotrexate as an intracellular marker for dihydrofolate reductase in L1210 cells.
A fluorescent derivative of methotrexate as an intracellular marker for dihydrofolate reductase in L1210 cells.
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甲氨蝶呤的荧光衍生物作为 L1210 细胞中二氢叶酸还原酶的细胞内标记物。
DOI：
10.1016/0003-9861(80)90402-6
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发表时间：
1980
影响因子：
3.9
通讯作者：
J. Whiteley
中科院分区：
文献类型：
--
作者：
G. Henderson;Andrea Russell;J. Whiteley
通过二氨基戊基连接基团与甲氨蝶呤偶联的异硫氰酸黄绿素（G.R. Gapski，J. M. Whiteley，J. I。雷德Cramer，G. B。亨德森、V. Neef和F. M. Huennekens，1975，J.Med.Chem.18，526-528）产生一种荧光化合物，它是从L1210鼠白血病细胞中纯化的二氢叶酸还原酶的强抑制剂（Ki= 60 nM）。荧光甲氨蝶呤衍生物优先采取的甲氨蝶呤耐药，而不是野生型L1210细胞培养中生长，并作为一个视觉标记的二氢叶酸还原酶（KD= 50 nM）在纯化和聚丙烯酰胺电泳。与二氢叶酸还原酶（通常是获得性细胞甲氨蝶呤抗性程度的指标）水平成比例的摄取缓慢发生，并通过与这些细胞转运甲氨蝶呤所用的载体介导系统不同的途径进行。
Fluorescein isothiocyanate coupled via a diaminopentyl-linking group to methotrexate (G.R. Gapski, J. M. Whiteley, J. I. Rader, P. L. Cramer, G. B. Henderson, V. Neef, and F. M. Huennekens, 1975,J. Med. Chem.18, 526–528) produces a fluorescent compound which is a strong inhibitor of dihydrofolate reductase (Ki= 60 nM) purified from L1210 murine leukemia cells. The fluorescent methotrexate derivative is preferentially taken up by methotrexate-resistant rather than wild-type L1210 cells grown in culture and acts as a visual marker for dihydrofolate reductase (KD= 50 nM) during both purification and polyacrylamide electrophoresis. Uptake, which is proportional to the level of dihydrofolate reductase (often an indicator of the degree of acquired cellular methotrexate resistance), occurs slowly and via a route that is distinct from the carrier-mediated system utilized by these cells to transport methotrexate.